Influence of RNA interference-induced silencing Smad4 gene on proliferation and migration of ECV304 cell line / 肿瘤

ABSTRACT

Objective: To investigate the effect of silencing Smad4 gene expression by using small interfering RNA (siRNA) on proliferation and migration of ECV304 cell line and explore the role of Smad4 in angiogenesis. Methods: Two siRNA sequences targeting Smad4 gene including siRNA1 and siRNA2 were designed and then synthetized. The two sequences of siRNA were transfected into ECV304 cells via lipofectamine mediation. The alteration of Smad4 mRNA and protein expression was examined by real-time RT-PCR and Western blot, respectively. The proliferation capacity of ECV304 cells was measured by cell cycle analysis and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) flow cytometry. The monostratal wound healing test was used to determine the migration capacity of ECV304 cells. Results: siRNA2 was successfully screened out of the two sequences. Transfection of siRNA2 significantly knocked down the Smad4 mRNA and protein expressions. The proliferation and migration capacity of ECV304 cells are significantly enhanced after transfection with siRNA2. Conclusions: Highly effective and specific siRNA targeting Smad4 gene can be successfully screened by RNA interference technique. Selective silencing of Smad 4 gene greatly promotes the proliferation and migration of ECV304 cells.